LOCUS       Exported                7555 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pMito.iGABASnFR.F102G.mRuby3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  AUTHORS   Marvin JS, Shimoda Y, Magloire V, Leite M, Kawashima T, Jensen TP, 
            Kolb I, Knott EL, Novak O, Podgorski K, Leidenheimer NJ, Rusakov DA,
            Ahrens MB, Kullmann DM, Looger LL
  TITLE     A genetically encoded fluorescent sensor for in vivo imaging of 
            GABA.
  JOURNAL   Nat Methods. 2019 Aug;16(8):763-770. doi: 10.1038/s41592-019-0471-2.
            Epub 2019 Jul 15.
  PUBMED    31308547
REFERENCE   2  (bases 1 to 7555)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7555
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             736..810
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1887..1908
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2011..2032)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3304..3321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3310..3534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3666..4254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3746..3765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4583..4630
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4655..4674
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4862..5650)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4973..4992)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5577..5596
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5685..6045)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5699..5837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5756..5775)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6002..6022
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(6105..6965)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6728..6747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6966..7070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7097..7552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7184..7203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7394..7415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct
      601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
      721 ttggggatat ccaccatgag cgtgctgacc cccctgctgc tgaggggcct gaccggcagc
      781 gccaggaggc tgcccgtgcc cagggccaag atccacagcc tgggcgaccc ccccgtggcc
      841 accagatctg aaagcatcaa tttcgtgagc tggggcggta gcacccagga tgcgcagaag
      901 caggcctggg ccgacccgtt cagcaaggcc agcggcatta ccgtggtcca ggatgggccc
      961 accgactacg gcaaactcaa ggccatggtc gaaagcggca acgtgcagtg ggacgtggtg
     1021 gatgtagagg ccgacttcgc cttgcgcgcc gccgccgaag gcctgctcga acccctggat
     1081 ttctcggtga tccagcgcga caagatcgac ccgcgcttcg tctccgacca tggcgtgggc
     1141 tcgttcctgg gctctttcgt gctcggctac aacgaaggca agctcggagc cagcaagccc
     1201 caggactgga ccgcgctgtt cgacaccaag acctaccccg gcaaacgcgc cctctacaaa
     1261 tggccgagcc ctggcgtgct cgaactggcc ctgctcgccg acggcgtacc ggccgacaag
     1321 ctctacccgc tggacctgga ccgcgccttc aagaaactcg acaccatcaa gaaagacatc
     1381 gtctggtggg gcggcggtgc acagtcgcag cagctgctgg cctccggcga agtcagcatg
     1441 ggccagttct ggaacggtcg catccacgcc ctgcaagaag acggcgcgcc agtgggcgtg
     1501 agctggaagc agaacctggt gatggccgac atcctcgtcg tgcctaaagg cacgaagaac
     1561 aaagccgccg cgatgaagtt cctggccagt gccagcagcg ccaaaggcca ggacgacttt
     1621 tccgccctga ccgcctatgc tccagtgaac atcgacagtg tgcagcgcct cgaccttgcg
     1681 caggtccgga tcaccgccga caagcagaag aacggcatca tggcgaactt caagatccgc
     1741 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1801 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgtgctgagc
     1861 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1921 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc
     1981 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2041 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2101 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2161 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2221 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
     2281 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2341 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2401 tggaacgcca acctggcacc gaacctgccg accgcctacg tcaaggatca gatcaccctc
     2461 gatttcgcct actgggccaa gaacggtccg gccattgcga cacggtggaa tgaatggcta
     2521 gtcaaactgc agggctccgg aaatgctgtg ggccaggaca cgcaggagcg tatggtgtct
     2581 aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc
     2641 aacggccacc aattcaaatg cacaggtgaa ggagaaggca ggccgtacga gggagtgcaa
     2701 accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc
     2761 acgtcgttca tgtatggcag ccgtactttt atcaagtacc cggcagacat tcctgatttc
     2821 tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt
     2881 ggagtcgtca ccgtcacgca ggacaccagc cttgaggatg gcgaactcgt ttacaacgtc
     2941 aaagtcagag gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt
     3001 tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg atacactgat
     3061 attgcactga aagttgatgg tggtggccat ctgcattgca atttcgtaac aacttacagg
     3121 tcaaaaaaga ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg
     3181 gaaaggatag aggaaagtga caatgaaacg tacgtagtac aacgcgaagt cgcagttgcc
     3241 aagtactcca atcttggtgg tggtatggat gaactttata agtaggcggc cgctcgagat
     3301 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3361 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3421 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3481 gggaggattg ggaagacaat agcaggcgtg ctggggatgc ggtgggctct atggcttctg
     3541 aggcggaaag aaccagtggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4081 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4381 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     4441 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     4501 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     4561 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     4621 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     4681 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     4741 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     4801 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     4861 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4921 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4981 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     5041 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     5101 cgacgagatc ctcgccgtcg ggcttgctcg ccttgagcct ggcgaacagt tcggctggcg
     5161 cgagcccctg atgctcttga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     5221 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     5281 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     5341 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     5401 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     5461 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5521 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     5581 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     5641 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     5701 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgagga ggcggcctcg
     5761 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     5821 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     5881 tgcttgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     5941 gactaattga gatgcttgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     6001 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc
     6061 aataaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7081 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7141 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7201 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7261 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7321 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7381 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7441 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7501 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacg
//
